Inhibition of tumor growth, invasion, and metastasis by targeting CD44 molecule

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1.Transfection of antisense CD44 (asCD44) into cancer cell : We created the stable transfectants by introducing a plasmid with asCD44 info a colorectal cancer cell line, LS174T. A mock transfectant (NEO) was also created.2.Invasion and migration of the asCD44 transfectants : There was no significant difference between asCD44 transfectants and NEO in in vitro assays of invasion and migration.3.Expression of agiogenic factors and MMPs : No significant difference was found in the expression of angiogenic factors and MMPs between asCD44 transfectants and the control.4.Subcutaneous implantation of the asCD44 transfectants : We inoculated the transfectants subcutaneously in nude mice. The tumor volume of the asCD44 transfectants was significantly smaller than that of NEO at day 3,7,14 after the inoculation. In a microscopic examination, the asCD44 transfectants were less angiogenic than NEO.5.Orthotopic implantation of the asCD44 transfectants : We injected the cells with MatrigelTM into submucosa of cecum in nude mice. No lymph node metastases were observed in mice with the asCD44 transfectants, although obvious lymph node metastases were observed in mice with NEO. Rate of the invasion of the asCD44 transfectants to lymph vessels was significantly lower than that of NEO.6.Adhesion of the transfectants to lymph nodes : The ability of the transfectants to adhere to human and mouse lymph nodes was examined in a Stamper-Woodruff assay. Adhesion of the asCD44 transfectants to the lymph nodes was significantly tower than that of NEO and parental cells. Hyaluronidase treatment of the parental cells reduced the attachment of the parenatal cells to the lymph nodes.In summary, the present study reveals that CD44 plays a crucial role in invasion of cancer cells into lymph vessels and lymph node metastases and that the interaction between CD44 and hyaluronate may be important in the lymph node metastases.